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What is the minimum DNA concentration requried for double digestion?

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#1 bochum



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Posted 13 April 2010 - 06:07 AM

i have less DNA and i like to digest the DNA using BglII and KpnI, product size is 400 and 8000kb. can any one suggest how much DNA in g, i have to use to see the positive clones from this digest.

Edited by bochum, 13 April 2010 - 06:08 AM.

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#2 phage434



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Posted 13 April 2010 - 09:43 AM

You need about 10 ng to see a band, so you need about 20x that much (8000/400) or 200 ng of DNA in your digestion, if you run the entire digestion. A standard digestion of 1 ug in 50 ul would be a good start, running 20 ul on a gel.

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